Severe signal loss on T2WI was observed in tumors of the CXCL12-NSPC group on day 42 but not in the tumors of the other groups ( Figure 2A). H&E staining indicated that this signal loss was attributable to intratumoral hemorrhage ( Figure 2B). As shown in Figure 2C (magnified

views check details of Figure 2B), an extensive area of hemorrhage (bright pink color on H&E staining) is clearly observed in the CXCL12-NSPC group. The hypointense areas were measured, and the ratios of the intratumoral hypointense areas were then calculated ( Figure 2D). The ratio of the hypointense area to that of the entire tumor region was significantly higher in the CXCL12-NSPC group than in the other groups (P < .001). The expression levels of CXCL12 and CXCR4 in the tumors of the four treatment groups were examined by immunohistochemistry (Figure 3).

Strong CXCL12 and CXCR4 expressions were detected in the CXCL12-NSPC group (Figure 3, CXCL12 and CXCR4). In addition, moderate CXCL12 and slight CXCR4 expressions were observed in the CXCL12-only group. The expression levels of CXCL12 and CXCR4 were either low or undetectable in the NSPC-only and sham groups. The grafted GFP-NSPCs 3-Methyladenine mw in the brains of animals in the CXCL12-NSPC and NSPC-only groups were identified by immunohistochemistry (Figure 4A, GFP). No GFP immunoreactivity was found in the CXCL12-only and sham groups, as expected, because GFP-NSPC transplantation was not employed in these groups. GFP+ cells were widespread in the tumors of the CXCL12-NSPC group, but only a few GFP+ cells were observed in the tumors of the NSPC-only group. A representative diagram of the distribution of GFP+ cells in the tumors of the CXCL12-NSPC group is shown in Figure 4B, in which each red dot represents two or three GFP+ cells. The number of GFP+ cells that had migrated toward tumor sites differed significantly between the CXCL12-NSPC (1159 ± 341

cells) and NSPC-only (45.7 ± 19.8 cells) groups (P < .01; Figure 4C). The grafted cells identified by GFP staining exhibited neuronal-like morphology with extended neurites (Figure 4A, magnified images from the CXCL12-NSPC and NSPC-only groups). Double labeling with NeuN (which is a neuronal marker) and GFP was employed to confirm the neuronal lineage of these GFP+ Tenofovir in vitro cells ( Figure 5A). GFP+/NeuN+ double staining demonstrated that ~ 80% of the GFP+ cells expressed NeuN in the tumors of the CXCL12-NSPC group (see Table 1). The number of GFP+/NeuN+ cells in the tumor regions ( Figure 5B) differed significantly between the CXCL12-NSPC (949 ± 258 cells) and NSPC-only (17.0 ± 14.6 cells) groups (P < .01; Figure 5B). Only a few NeuN+ cells were found in the CXCL12-only and sham groups (data not shown). The targeted migration of stem cells is essential for the direct repair of injured tissues.